Purification of galectin-3 from ovine placenta: developmentally regulated expression and immunological relevance.
Galectins, beta-galactoside-binding lectins, are extensively distributed in the animal kingdom and share some basic molecular properties. Galectin-3, a member of this family, is generally associated with differentiation, morphogenesis, and metastasis. In this study, galectin-3 was isolated from ovine placental cotyledons round the middle of the gestation period by lactose extraction followed by affinity chromatography on lactosyl-agarose, and separated from galectin-1 by size exclusion chromatography on a Superose 12 column. Under native conditions this lectin behaved as a monomer with an apparent molecular weight of approximately 29,000 and an isoelectric point of 9.0. The partial amino acid sequence of the peptides obtained by tryptic digestion of this protein followed by HPLC separation showed striking homology with other members of the galectin-3 subfamily. Furthermore, ovine placental galectin-3 exhibited specific mitogenic activity toward rat spleen mononuclear cells. Besides, this protein strongly reacted with a rabbit antiserum raised against a chicken galectin. Results obtained by Western blot analysis showed that its expression was greatly decreased in term placenta with respect to the middle of the gestation period, suggesting a regulated expression throughout development.
Iglesias
M M
MM
Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Facultad de Farmacia y Bioquímica, Junín 956, 1113 Buenos Aires, Argentina.
Rabinovich
G A
GA
Ambrosio
A L
AL
Castagna
L F
LF
Sotomayor
C E
CE
Wolfenstein-Todel
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002455:Cell Division / Q000187:drug effects; D003208:Concanavalin A / Q000494:pharmacology; D005260:Female; D037502:Galectin 3; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007526:Isoelectric Point; D008970:Molecular Weight; D010920:Placenta / Q000737:chemistry* / Q000276:immunology; D010929:Placentation; D011247:Pregnancy; D011817:Rabbits; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012756:Sheep
D000943:Antigens, Differentiation; D037502:Galectin 3; D003208:Concanavalin A